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In Vivo Tagging and Characterization of S‐Glutathionylated Proteins by a Chemoenzymatic Method
Author(s) -
Chiang BingYu,
Chou ChiChi,
Hsieh FuTan,
Gao Shijay,
Lin Jason ChingYao,
Lin ShengHuang,
Chen TzeChieh,
Khoo KayHooi,
Lin ChunHung
Publication year - 2012
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201200321
Subject(s) - cysteine , glutathione , biochemistry , chemistry , biotin , s nitrosylation , proteomics , biotinylation , enzyme , gene
Glutathionylspermidine synthetase (GspS) was introduced into human cells to label endogenous glutathione (GSH) with biotinyl spermine (biotin‐spm) for the identification of S‐thiolated proteins. The proteins carrying Gspm‐biotin can be detected by immunoblotting (A in figure). Enrichment by using the biotin tag enabled subsequent mass‐spectrometry‐based proteomic analysis (B) for site‐specific identification of glutathionylated sites.